Effect of circ_0084043 silence upon human renal tubular epithelial cell injury induced by high glucose
ObjectiveTo explore the effect of silencing circular RNA_0084043（circ_0084043）on the inflammatory responses， oxidative stress and apoptosis of human renal tubular epithelial cells induced by high glucose and its targeted regulation of miR-142-5p.MethodsRenal tubular epithelial cells HK-2 were divide...
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| Main Authors: | , |
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| Format: | Article |
| Language: | zho |
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Editorial Department of Journal of Clinical Nephrology
2022-09-01
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| Series: | Linchuang shenzangbing zazhi |
| Subjects: | |
| Online Access: | http://www.lcszb.com/thesisDetails#10.3969/j.issn.1671-2390.2022.09.010 |
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| Summary: | ObjectiveTo explore the effect of silencing circular RNA_0084043（circ_0084043）on the inflammatory responses， oxidative stress and apoptosis of human renal tubular epithelial cells induced by high glucose and its targeted regulation of miR-142-5p.MethodsRenal tubular epithelial cells HK-2 were divided into the groups of Con， HG， HG+si-NC， HG+si-circ_0084043， HG+miR-NC， HG+miR-142-5p， HG+si-circ_0084043+anti-miR-NC and HG+si-circ_0084043+anti-miR-142-5p. Quantitative real-time polymerase chain reaction （qRT-PCR） was utilized for detecting the expression levels of circ_0084043 and miR-142-5p according to the random number table Enzyme-linked immunosorbent assay （ELISA） was utilized for detecting the levels of tumor necrosis factor-alpha （TNF-α） and interleukin-8 （IL-8）. The levels of malondialedhyde （MDA） and superoxide dismutase （SOD） were detected. Flow cytometry was employed for detecting apoptotic rate. Dual luciferase reporter gene was utilized for detecting the targeting relationship between circ_0084043 and miR-142-5p and Western blot for detecting the expressions of Bcl-2 and Bax protein.ResultsAs compared with Con group， the expression level of circ_0084043 spiked in HG group ［（1.00±0.04） <italic>vs</italic> （3.57±0.49）］（<italic>P</italic><0.05） while the expression level of miR-142-5p declined ［（1.00±0.05） <italic>vs</italic> （0.34±0.06）］ （<italic>P</italic><0.05）； the levels of TNF-α， IL-8 and MDA ［（18.43±2.06） <italic>vs</italic> （49.55±2.65） mg/mL； （11.74±0.83） <italic>vs </italic>（27.62±2.25） mg/mL； （0.23±0.08） <italic>vs</italic> （2.86±0.37） nmol/mg］（<italic>P</italic><0.05）， apoptotic rate ［（6.74±0.95）% <italic>vs</italic> （23.15±2.49）%］ and protein level of Bax increased （<italic>P</italic><0.05） while activity of SOD decreased ［（40.05±3.07） <italic>vs</italic> （17.35±1.54） U/mg］ （<italic>P</italic><0.05） and protein level of Bcl-2 dropped （<italic>P</italic><0.05）. Compared with HG/HG+si-NC group， the levels of TNF-α， IL-8 and MDA declined in HG+si-circ_0084043 group ［（49.55±2.65）/（48.42±4.04） <italic>vs</italic> （34.90±4.95） mg/mL； （27.62±2.25）/（27.76±3.53） <italic>vs </italic>（19.14±1.02） mg/mL； （2.86±0.37）/（2.81±0.43）<italic>vs</italic>（0.89±0.12） nmol/mg］（<italic>P</italic><0.05） and apoptotic rate ［（23.15±2.49）/（23.73±3.28）% <italic>vs</italic> （14.47±0.93）%］ and protein level of Bax declined （<italic>P</italic><0.05） while the activity of SOD ［（17.35±1.54）/（17.40±0.63） <italic>vs</italic> （28.54±2.56） U/mg］ （<italic>P</italic><0.05） and protein level of Bcl-2 rose （<italic>P</italic><0.05）. And circ_0084043 could target miR-142-5p. As compared with HG+miR-NC group， the levels of TNF-α， IL-8 and MDA declined in HG+miR-142-5p group ［（48.78±4.56） <italic>vs</italic> （35.18±3.93） mg/mL； （27.98±1.64） <italic>vs</italic> （18.31±1.11） mg/mL； （2.81±0.32） <italic>vs </italic>（0.96±0.08） nmol/mg］（<italic>P</italic><0.05） and apoptotic rate ［（22.27±2.68） % <italic>vs</italic> （16.01±0.48）%］ and protein level of Bax dropped （<italic>P</italic><0.05） while activity of SOD ［（17.40±1.09） <italic>vs</italic> （25.22±1.67） U/mg］（<italic>P</italic><0.05） and protein level of Bcl-2 rose （<italic>P</italic><0.05）. As compared with HG+si-circ_0084043+anti-miR-NC group， the levels of TNF-α， IL-8 and MDA rose in HG+si-circ_0084043+anti-miR-142-5p group ［（34.74±1.51） <italic>vs </italic>（46.31±2.65） mg/mL； （19.03±0.66） <italic>vs</italic> （23.07±1.19） mg/mL； （0.89±0.06） <italic>vs</italic> （1.43±0.11） nmol/mg］ （<italic>P</italic><0.05） and apoptotic rate ［（14.69±0.49）% <italic>vs</italic> （18.37±0.89）%］ and protein level of Bax spiked （<italic>P</italic><0.05） while the activity of SOD ［（28.38±1.03） <italic>vs</italic> （19.56±0.83） U/mg］（<italic>P</italic><0.05） and protein level of Bcl-2 declined （<italic>P</italic><0.05）.ConclusionSilencing circ_0084043 could inhibit cell inflammatory responses， oxidative stress and apoptosis through negatively regulating miR-142-5p， thereby blunting the damage of human renal tubular epithelial cells induced by high glucose. |
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| ISSN: | 1671-2390 |